Two stages of decomposition are the key step of pyrolysis since they took up half or more of the reaction time; melting step consumed another half of reaction time in experiments when raw materials were heated up from ambient temperatures; and coke-like deposition appeared as a result of decomposition completion.
In order to achieve homogeneous products of n-type Ti1−xZrxNiSn, Ti1−xHfxNiSn, and corresponding Sn/Sb substituted alloys, the synthesis conditions were tuned by an optimized arc melting technique, an intermediate high frequency melting step, long time ball milling and a multi-step hot pressing technique.
b Micromolding melting step.
Although the amplification reaction of NASBA is isothermal, a single melting step prior to the amplification reaction is required to allow annealing of the primers to the target, which is not required in LAMP [35, 36].
For the melting step, the temperature was raised from 75°C to 97°C with a ramp rate of 0.02°C/s and 25 acquisitions/°C.
Samples were subjected to a melting step of 95°C for 10 min followed by 40 cycles of 15 s at 95°C and 1 min at 60°C.
A melting step was then performed with continuous fluorescence acquisition starting at 60°C with a rate of 1°C/5 s up to 99°C to determine the amplification specificity.
After the thermocycling reaction the melting step was performed with slow heating, starting at 55°C and with a rate of 0.5°C per 10 s, up to 95°C, with continuous measurement of fluorescence, allowing detection of possible non-specific products.
The melting step was performed using 30 acquisitions per °C.
Amplicon integrity was evaluated by fluorescence reading, melting step, and 2% agarose ethidium bromide gels.
Before the high resolution melting step the products were heated to 95°C for 1 minute.
